Collagen1和Collagen3在牦牛肺纤维化组织中的表达研究

为探讨Collagen1和Collagen3在牦牛肺纤维化组织中的表达,阐明Collagen1和Collagen3在牦牛肺纤维化进程中发挥的作用,采集牦牛正常肺组织和发生纤维化的肺组织,将其分为对照组和试验组,通过HE染色、Masson染色及透射电镜观察肺超微结构的病理变化及纤维化状态;利用实时荧光定量PCR(qRT-PCR)、蛋白免疫印迹(WB)、免疫组化和免疫荧光染色法检测和定位Collagen1、Collagen3基因和蛋白的表达情况。结果显示,试照组牦牛肺组织结构完整,肺泡隔无增厚,支气管管腔和肺泡腔内均无炎性渗出物;试验组肺组织可见片状坏死,胞核碎裂溶解,重度出血,大范围肺水肿。Collagen1在试验组中的表达量高于对照组,而Collagen3在对照组中的表达量高于试验组。在试验组中,Collagen1和Collagen3蛋白大量增生,在肺泡隔中有大量分布,其余分布位置与肺对照组基本一致,但表达都高于对照组。综上所述,Collagen1和Collagen3在牦牛发生肺纤维化的过程中发挥着重要作用,Collagen1和Collagen3的相对表达量在诊断牦牛纤维化肺炎中具有一定的参考价值。

Low-molecular-weight fucoidan inhibits the proliferation of melanoma via Bcl-2 phosphorylation and PTEN/AKT pathway

Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance.

Production and Characterization of Recombinant Rat Non-Collagen Domain of α3 Chain of Type IV Collagen α3 (IV) NC1 Antigen

The glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney that causes kidney diseases. The reason of glomerulonephritis disease is to deposit the anti-GBM auto antibody in the glomerular basement membrane. The type IV collagen is the main component of glomerular basement membrane that has α3 chain of type (IV) collagen of non-collagenous domain which contains N-terminal 7S domain, a triple helical collagenous domain and C-terminal non-collagenous glomerular domain (NC1). The amino terminal of α3 (IV) NC1 that induces the Experimental Autoimmuno Glomerulonephritis (EAG) in rat model has been identified. The recombinant rat α3 (IV) NC1 antigen has nine amino acid spans that are consistent with antibody or T cell epitope that induces in EAG. The research is carried out on the recombinant rat α3 (IV) NC1 production, purification, quantification, and characterization. The circulation of anti-GBM antibody in glomerular basement membrane can be measured by the ELISA assay. In addition, the recombinant rat antigen is secreted in HEK293 cell supernatant that is purified by Anti-FLAG M2 monoclonal IgG antibody affinity column and characterized and quantified by SDS-PAGE gel electrophoresis and Western blotting techniques.

NC1-peptide derived from collagenα3(IV)chain is a blood-tissue barrier regulator:lesson from the testis

Collagenα3(IV)chains are one of the major constituent components of the basement membrane in the mammalian testis.Studies have shown that biologically active fragments,such as noncollagenase domain(NC1)-peptide,can be released from the C-terminal region of collagenα3(IV)chains,possibly through the proteolytic action of metalloproteinase 9(MMP9).NC1-peptide was shown to promote blood-testis barrier(BTB)remodeling and fully developed spermatid(e.g.,sperm)release from the seminiferous epithelium because this bioactive peptide was capable of perturbing the organization of both actin-and microtubule(MT)-based cytoskeletons at the Sertoli cell-cell and also Sertoli-spermatid interface,the ultrastructure known as the basal ectoplasmic specialization(ES)and apical ES,respectively.More importantly,recent studies have shown that this NC1-peptide-induced effects on cytoskeletal organization in the testis are mediated through an activation of mammalian target of rapamycin complex 1/ribosomal protein S6/transforming retrovirus Akt1/2 protein(mTORC1/rpS6/Akt1/2)signaling cascade,involving an activation of cell division control protein 42 homolog(Cdc42)GTPase,but not Ras homolog family member A GTPase(RhoA),and the participation of end-binding protein 1(EB1),a microtubule plus(+)end tracking protein(+TIP),downstream.Herein,we critically evaluate these findings,providing a critical discussion by which the basement membrane modulates spermatogenesis through one of its locally generated regulatory peptides in the testis.

Derived vascular endothelial cells induced by mucoepidermoid carcinoma cells:3-dimensional collagen matrix model

Mucoepidermoid carcinoma undergoes uniquely vigorous angiogenic and neovascularization processes,possibly due to proliferation of vascular endothelial cells(ECs) induced by mucoepidermoid carcinoma cells(MCCs) in their three-dimensional(3D) microenvironment.To date,no studies have dealt with tumor cells and vascular ECs from the same origin of mucoepidermoid carcinoma using the in vitro 3D microenvironment model.In this context,the current research aims to observe neovascularization with mucoepidermoid carcinoma microvascular ECs(MCMECs) conditioned by the microenvironment in the 3D collagen matrix model.We observed the growth of MCMECs purified by immunomagnetic beads and induced by MCCs,and characteristics of tubule-like structures(TLSs) formed by induced MCMECs or non-induced MCMECs.The assessment parameters involved the growth curve,the length,the outer and inner diameters,and the wall thickness of the TLSs,and the cell cycle.Results showed that MCCs induced formation of the TLSs in the 3D collagen matrix model.A statistically significant difference was noted regarding the count of TLSs between the control group and the induction group on the 4th day of culture(t=5.00,P=0.001).The outer and inner diameters(t1=5.549,P1=0.000;t2=10.663,P2=0.000) and lengths(t=18.035,P=0.000) of the TLSs in the induction group were statistically significant larger than those in the control group.The TLSs were formed at the earlier time in the induction group compared with the control group.It is concluded that MCCs promote growth and migration of MCMECs,and formation of the TLSs.The 3D collagen matrix model with MCMECs induced by MCCs in the current research may be a favorable choice for research on pro-angiogenic factors in progression of mucoepidermoid carcinoma.